Abstract：
In this study, we report the innovative application of whole-cell patch-clamp electrophysiology in assessing broad-spectrum neutralisation by three different antivenoms, of venoms from the medically significant scorpion genus Centruroides. Envenomations by as many as 21 species from the Centruroides genus result in up to 300,000 envenomations per year in Mexico, which poses significant and potentially life-threatening pathophysiology. We first evaluated the in vitro manifestation of envenomation against two human voltage-gated sodium (hNaV) channel subtypes: hNaV1.4 and hNaV1.5, which are primarily expressed in skeletal muscles and cardiomyocytes, respectively. The neutralisation of venom activity was then characterised for three different antivenoms using a direct competition model against the more potent target, hNaV1.4. While broad-spectrum neutralisation was identified, variation in neutralisation arose for Centruroides elegans, C. limpidus, C. noxius and C. suffusus venoms, despite the presence of a number of these venoms within the immunising mixture. This raises questions regarding the truly \"broad\" neutralisation capacity of the antivenoms. This study not only extends previous validation of the in vitro investigation of antivenom efficacy utilising the whole-cell patch-clamp technique but also underscores the potential of this animal-free model in exploring cross-reactivity, experimental scalability, and most importantly, informing clinical management practices regarding the administration of antivenom in Mexico.
摘要：
在这项研究中,我们报告了全细胞膜片钳电生理学在评估三种不同抗蛇毒血清广谱中和方面的创新应用，来自医学上重要的蝎子属的毒液。来自Centruroides属的多达21种的毒素在墨西哥每年导致多达300,000种毒素，这构成了重要的和潜在的威胁生命的病理生理学。我们首先评估了两种人电压门控钠（hNaV）通道亚型的体外表现：hNaV1.4和hNaV1.5，它们主要在骨骼肌和心肌细胞中表达，分别。然后使用针对更有效靶标的直接竞争模型，对三种不同的抗蛇毒血清进行了毒液活性的中和表征，hNaV1.4.虽然发现了广谱中和，秀丽隐杆线虫出现中和变化，C.limpidus,C.Noxius和C.充满毒液，尽管免疫混合物中存在许多这些毒液。这引发了有关抗蛇毒血清真正的“广泛”中和能力的疑问。这项研究不仅扩展了先前使用全细胞膜片钳技术对抗蛇毒血清功效的体外研究的验证，而且还强调了这种无动物模型在探索交叉反应性方面的潜力。实验性的可扩展性，最重要的是,告知墨西哥抗蛇毒血清管理的临床管理实践。
